Rohwedel J, Sehlmeyer U, Shan J, Meister A, Wobus A M
Institut für pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.
Cell Biol Int. 1996 Aug;20(8):579-87. doi: 10.1006/cbir.1996.0076.
Embryonic germ (EG) cells of line EG-1 derived from mouse primordial germ cells were investigated for their in vitro differentiation capacity. By cultivation as embryo-like aggregates EG-1 cells differentiated into cardiac, skeletal muscle and neuronal cells accompanied by the expression of tissue-specific genes and proteins as shown by RT-PCR analysis and indirect immunofluorescence. In comparison to embryonic stem (ES) cells of line D3 the efficiency of differentiation into cardiac and muscle cells was comparatively low, whereas spontaneous neuronal differentiation was more efficient than in D3 cells. Furthermore, the distribution of cell cycle phases as a parameter for the differentiation state was analysed in undifferentiated EG cells and ES cells and compared to data obtained for embryonic carcinoma (EC) cells of line P19 and differentiated, epithelioid EPI-7 cells. Flow cytometric analysis revealed similar cell cycle phase distributions in EG, EC and ES cells. In contrast, the somatic differentiated EPI-7 cells showed a longer G1-phase and shorter S- and G2/M-phases. Together, our results demonstrate that the differentiation state and capacity of EG cells in vitro resemble that of totipotent ES cells.
对源自小鼠原始生殖细胞的EG-1系胚胎生殖(EG)细胞的体外分化能力进行了研究。通过培养成胚胎样聚集体,EG-1细胞分化为心脏、骨骼肌和神经元细胞,同时伴随着组织特异性基因和蛋白质的表达,逆转录-聚合酶链反应(RT-PCR)分析和间接免疫荧光显示了这一点。与D3系胚胎干细胞(ES细胞)相比,EG-1细胞分化为心脏和肌肉细胞的效率相对较低,而自发神经元分化比D3细胞更有效。此外,分析了未分化的EG细胞和ES细胞中作为分化状态参数的细胞周期阶段分布,并与P19系胚胎癌细胞(EC细胞)和分化的上皮样EPI-7细胞的数据进行了比较。流式细胞术分析显示EG、EC和ES细胞具有相似的细胞周期阶段分布。相比之下,体细胞分化的EPI-7细胞显示出更长的G1期和更短的S期及G2/M期。总之,我们的结果表明EG细胞在体外的分化状态和能力类似于全能ES细胞。